Analytical purification always requires some kind of chromatography. You can develop a sort of affinity based separation if your compound has affinity towards any liquid or solid surface you can use or exhibits differential solubility.
Extraction with distill water and put into the magnetic water bath mixer
(600 rpm, 90°C), after 4 h, the suspension centrifuged, the extract sediments twice with the same method. Then collect supernatants of the three centrifuge merged and concentrated at 50°C by evaporation under reduced pressure. Add three volumes of ethanol precipitation of polysaccharide at 4C overnight, then precipitation will obtain by centrifugation (5000 rpm, 10 min) repeatedly washed with ethanol. Repeat extraction thrice
gudluck
:)
ThanKs Robin Joshi, I have already done the precipitation with ethanol. And i think it is a mixture of polysaccharides. I had extracted with water only and lyophilised and precipitated. I think proteins may also precipitate while doing this. After the precipitation can I directly go to Characterization ?
Yes go for characterisation. You can also go for their biocativities such as anticancer etc.
Thanks Robin Joshi Is it enough pure for taking NMR after precipitation..?
and what about glycoproteins..? they may also prectipitate.. right..?
Before NMR use TLC analysis with a developing reagent (ethyl acetate: pyridine:
glacial acetic acid: water in a 5:5:1:3 volume ratio for saccrides. This may be help you out.
Thanks Joshi, TLC is doing to know about the purity right..? I know that the crude precipitate of polysaccharides needs purification.. TLC is enough..?
TLC is only for detection. For charcatristion go for LC-MS and NMR. I will send reference paper tomorrow.
gudngt
you can use molecular cut off filter.. or there are a range of other resins.. you can also look at electrophoresis.
Thanks Haroon Iqbal Did you mean Dialysis membrane..? What could be the cutt of filter I can use..?
http://www.millipore.com/userguides/files/centrifugal/$file/PR03520TR_RevA_English.pdf
you can also try size exclusion chromatography, using resins http://www.gelifesciences.com/webapp/wcs/stores/servlet/catalog/en/GELifeSciences/brands/sephadex/
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